肾消解毒通络方对糖尿病肾病金黄地鼠肾脏AIM2介导细胞焦亡的影响

Effects of Shenxiao Jiedu Tongluo Recipe on AIM2-mediated pyroptosis of renal cells in a golden hamster model of diabetic nephropathy

目的探讨肾消解毒通络方对糖尿病肾病金黄地鼠肾脏AIM2介导细胞焦亡的影响及其作用机制。方法50只SPF级雄性金黄地鼠随机分为对照组和造模组,造模组采用高糖高脂喂养联合腹腔注射STZ构建糖尿病肾病模型,建模成功后随机分为模型组、恩格列净组(10 mg/kg)和肾消解毒通络方低、高剂量组(12.8、25.6 g/kg),灌胃给药8周。全自动生化分析仪检测空腹血糖、24 h-UTP和血清TC、LDL-C、Scr、Sur水平,生化法检测血清SOD活性和MDA水平,ELISA法检测血清IL-1β、IL-18、TNF-α水平,HE和PAS染色观察金黄地鼠肾组织病理形态学变化,免疫荧光或免疫组化法检测肾组织ROS、γH2AX蛋白表达,Western blot法及免疫组化法检测肾组织AIM2、caspase-1、GSDMD蛋白表达。结果与对照组比较,模型组金黄地鼠肾小球出现萎缩,肾小球囊腔增大、系膜区增宽,肾小管扩张、水肿伴有坏死,空腹血糖、24 h-UTP、血清TC、LDL-C、Scr、Sur、IL-1β、IL-18、TNF-α、MDA水平和肾组织ROS、γH2AX、AIM2、caspase-1、GSDMD蛋白表达升高(P<0.01),血清SOD活性降低(P<0.01);与模型组比较,肾消解毒通络方高剂量组和恩格列净组肾组织病理学得到改善,空腹血糖、24 h-UTP、血清TC、LDL-C、Scr、Sur、IL-1β、IL-18、TNF-α、MDA水平和肾组织ROS、γH2AX、AIM2、caspase-1、GSDMD蛋白表达降低(P<0.05,P<0.01),血清SOD活性升高(P<0.01)。结论肾消解毒通络方可通过抑制金黄地鼠ROS-dsDNA-AIM2信号通路表达,降低肾组织ROS水平,减少肾脏固有细胞DNA损伤,减少AIM2炎症小体合成,从而抑制AIM2介导的细胞焦亡,减轻肾脏炎症损伤。

AIM To investigate the effects and mechanism of Shenxiao Jiedu Tongluo Recipe on renal AIM 2-mediated pyroptosis of a golden hamster model of diabetic nephropathy(DN).METHODS Fifty male golden hamsters of SPF grade were randomly divided into the control group and the model group.The golden hamsters of the model group successfully developed into DN models by feeding of high glucose and high fat diet and intraperitoneal injection of STZ were further randomly assigned into the model group,the enagliflozin group(10 mg/kg),and the low-dose and the high-dose Shenxiao Jiedu Tongluo Recipe groups(12.8,25.6 g/kg)for 8 weeks gavage of the corresponding administration.The golden hamsters had their levels of fasting blood glucose,24 h-UTP,serum TC,LDL-C,Scr,and Sur detected by automatic biochemical analyzer;their serum SOD activity and MDA level detected by biochemical method;their serum levels of IL-1β,IL-18,and TNF-αdetected by ELISA method;their pathomorphological changes of kidney tissue observed by HE and PAS staining;their protein expressions of ROS andγH2AX detected by immunofluorescence or immunohistochemistry;and their renal protein expressions of AIM 2,caspase-1 and GSDMD detected by Western blot and immunohistochemistry.RESULTS Compared with the control group,the model group showed atrophic glomeruli;enlarged glomerular capsule cavity;mesangial expansion;edema and necrosis in the dilated renal tubules;increased levels of fasting blood glucose,24 h-UTP,serum TC,LDL-C,Scr,Sur,IL-1β,IL-18,TNF-α,MDA and renal protein expressions of ROS,γH2AX,AIM2,caspase-1,GSDMD(P<0.01);and decreased serum SOD activity(P<0.01).Compared with the model group,the high-dose Shenxiao Jiedu Tongluo Recipe group and the enagliflozin group displayed improved renal histopathology,decreased levels of 24 h-UTP,serum TC,LDL-C,Scr,Sur,IL-1β,IL-18,TNF-α,MDA and renal protein expressions of ROS,γH2AX,AIM2,caspase-1,GSDMD(P<0.05,P<0.01);and increased serum SOD activity(P<0.01).CONCLUSION Shenxiao Jiedu Tongluo Recipe can inhibit AIM 2-mediated cell death and alleviate renal inflammatory damage in golden hamsters by inhibiting their expression of ROS-dsDNA-AIM 2 signal pathway to attain reduction of their renal ROS level,DNA damage of renal intrinsic cells,and synthesis of AIM 2 inflammatory corpuscles as well.